Anik Girard-Globa

Postprandial cholesteryl ester transfer and high density lipoprotein composition in normotriglyceridemic non-insulin-dependent diabetic patients.

Altered postprandial HDL metabolism is a possible cause of defective reverse cholesterol transport and increased cardiovascular risk in diabetic patients with a normal fasting lipoprotein profile. Ten normolipidemic, normoponderal non-insulin dependent diabetes mellitus (NIDDM) patients and seven controls received a 980 kcal meal containing 78 g lipids with 100 000 IU vitamin A. Chylomicron clearance was not different, but area under the curve (AUC) for retinyl palmitate in chylimicron-free serum (remnant clearance) was greater in patients (P < 0.02). LCAT activity increased postprandially to the same extent in both groups. In control subjects, cholesteryl ester transfer protein (CETP) activity (CETA) also increased by 20% (P < 0.01 at 6 h) in parallel with a 20% decrease in HDL2-CE (r = -0.55, P = 0.009). In NIDDM patients, on the contrary, CETA which was 35% higher in the fasting state (P < 0.005), decreased postprandially yet HDL2-CE remained unchanged. Postprandial HDL3 of controls were enriched with phospholipid (PL) (30.3 +/- 2.6% at 6 h) with respect to fasting (25.6 +/- 2.5%, P < 0.01) and to NIDDM-HDL3 (25.8 +/- 1.7% at 6 h, P < 0.01). These results show that variation in plasma CETA has little impact on HDL2-CE in NIDDH subjects. They support the concept that, in controls, the combined enrichment of HDL3 with PL, increased LCAT and CETA create the conditions for stimulation of cell cholesterol efflux and CE transfer to apo B lipoproteins. In NIDDM, because of the lesser HDL3 enrichment with PL and of the inverse trend of CETA, these conditions fail to occur, depriving the patients of a potentially efficient mechanism of unesterified cholesterol (UC) clearance, despite their strictly normal preprandial profile.

Modulation of low density lipoprotein subclasses by alimentary lipemia in control and normotriglyceridemic non-insulin-dependent diabetic subjects.

Conventional factors do not fully account for the increased cardiovascular risk in NIDDM but, because of the underlying disorders in lipid metabolism, the postprandial state can be expected to induce temporary changes of a potentially atherogenic nature. The response to a 1000-kcal meal (70% lipid; 100,000 IU vitamin A) over 8 h was compared in 10 normoponderal, normotriglyceridemic NIDDM male patients and 12 controls. In patients lipolysis was normal, but remnant clearance was delayed (P < 0.02) and apo E concentrations were lower. LDL-C decreased postprandially, more in patients (P < 0.05), while LDL-PL accumulated in controls but not in patients. As a result UC:PL decreased in controls (P < 0.05) not in patients. The distribution of LDL subclasses shifted towards large particles in controls (LDL-I, 42%; LDL-II, 50%; LDL-III, 7.6% at 6 h) and smaller ones in patients (LDL-I, 29%; LDL-II, 56%; LDL-III, 16% at 6 h). In controls only, the percentage of LDL-III correlated negatively with apo E (r = -0.97, P < 0.001) suggesting that apo E promotes removal of light particles before they reach LDL-III and may be a limiting factor in patients. We conclude that the postprandial state is potentially more atherogenic in normoponderal, normotriglyceridemic patients than in controls: remnant clearance is delayed, the UC:PL ratio of LDL fails to decrease postprandially as it does in controls, limiting the acceptor capacity of LDL for UC, and the distribution of LDL subclasses is shifted towards a more atherogenic profile.

Oleic acid-rich increase increase the capacity of postprandial serum to promote cholesterol efflux from Fu5AH cells

Abstract Cell cholesterol efflux to serum is stimulated after an oral fat load. The impact of meal fatty acid composition was explored by measure of serum promoted cholesterol efflux from Fu5AH cells after ingestion of 4 different fats: sunflower (Sf), oleic-sunflower (Ol), a mixed oil (Mx), and beef tallow (Bt). High density lipoprotein (HDL)2 and HDL3 were isolated and analyzed. Cholesterol efflux increased regularly after Ol (P

Postprandial concentrations and distribution of apo C-III in type 2 diabetic patients. Effect of bezafibrate treatment

Apo C-III plays a key role in the metabolism of triglyceride-rich lipoproteins. It has recently been implicated as a potential determinant of the triglyceride (TG) lowering effect of fibrates, which down-regulate its expression. This hypothesis has been explored in ten moderately hypertriglyceridemic (TG 4.50+/-2.40 mmol/l) male type 2 diabetic patients tested with a lipid load before and after 4 weeks of treatment with 400 mg bezafibrate daily. Treatment lowered apo C-III concentrations by 20%, mainly in VLDL. Postprandially, apo C-III was transferred to chylomicrons in proportion to their TG content exclusively from HDL. VLDL retained their apo C-III and the apo C-III:TG ratio decreased as TG contents increased. At the end of the absorptive period (8 h) HDL did not recover the totality of their apo C-III (net loss 19 and 28% respectively before and after treatment, P<0.0001 for time effect). Bezafibrate lowered apo E by 33% (P<0.03). The apo C-III:apo E ratio did not vary significantly under treatment but underwent a postprandial decrease: 13% before and 18% (P=0.01) after treatment. These results indicate that repression of apo C-III expression and lowering of the apo C-III:E ratio are not likely mechanisms for the lipid-lowering effects of fibrates in type 2 diabetic patients. The potent effects on postprandial lipemia are suggestive of an apo C-III-independent stimulation of lipolysis.

Distribution of HDL2 and HDL3 in a random population of healthy French males and females —evaluation by a two-step precipitation procedure

A rapid and reliable method for the determination of HDL2 and HDL3 cholesterol is described. The Apo B containing fractions (VLDL, IDL, LDL) were precipitated by addition of dextran sulfate (Mr 500,000) to 2 mmol/l final concentration followed by MgCl2 to a final concentration of 0.05 mol/l. The supernatant, was brought to 6 mmol/l dextran sulfate and 0.250 mol/l MgCl2 to precipitate HDL2. Cholesterol determination on total serum and both supernatants yielded the concentrations of Apo B-associated cholesterol, total HDL, HDL2 and HDL3 cholesterol. The application of this technique to a random population of healthy French people gave HDL-cholesterol values of 1.35 and 1.54 mmol.l-1, respectively, in 93 males and 95 females (p less than 0.001). All of the difference was attributable to HDL2 (0.43 vs 0.65 mmol.l-1, p less than 0.001) while HDL3 were almost identical at 0.92 and 0.91 nmol.l-1. These values are in good agreement with previously reported figures for French individuals, but markedly higher in males than values reported from North America.

Regulation of hepatic synthesis of proteins by the chronology of protein ingestion

Circadian variations in liver protein synthesis were were assessed in control rats fed a mixed 10% protein diet and in rats fed proteins as a separate meal either at 09:00 (SF 09) or at 21:00 (SF 21) and provided with a protein-free diet ad libitum. Protein synthesis was measured by incorporation of labelled leucine over a short period of time (15 min) at time-points regularly spaced over 24 h. In controls, the circadian variations observed were of moderate amplitude (from 2.75 mg/h per g at 09:00 to 5.77 mg/h per g at 06:00) correlated with increased protein and RNA contents of the liver. In separately fed animals ingestion of the protein meal triggered a 300% increase in protein synthesis within 1 h while the feeding pattern was unaltered. In the SF 09 group, high synthetic activity was not followed by an increase of hepatic protein content while hepatic urea concentrations were sharply increased and glucogenic amino acid pools were greatly depleted. It is suggested that the high influx of amino acids consecutive to the absorption of the dietary proteins is the key factor stimulating protein synthesis, while synchronisation with the energetic metabolism controls the degree of degradation. The possible involvement of variations in the insulin to glucagon ratio is discussed.

Induction of multiple forms of tyrosine aminotransferase by amino acid mixtures of different compositions

Hepatic tyrosine aminotransferase (EC 2.6.1.5) was induced in rats by intubation of amino acid mixtures (complete or tryptophan-free). Enzyme activity was increased 4-fold by the complete mixture and 8-fold by the tryptophan-free mixture. The enzyme was analyzed by chromatography on CM-Sephadex. Chromatographic patterns were characteristic of the type of inducer rather than of the chronology of the induction cycle: after induction by the complete amino acid mixture the three forms of the enzyme were equally increased whereas after induction by the tryptophan-free mixture Form I was preferentially increased.

Lipoproteins and Apoproteins of Fetal and Newborn Piglets

Serum lipids, very low in the fetus and at birth, increased rapidly after the first feeding. Cholesterol was mainly in the free form. Esterified cholesterol rose rapidly, but free cholesterol remained in high proportions (30%) until after weaning. Very high phospholipid concentrations show that triglycerides are not the only major product of esterification and suggest a possible use for energetic purposes. Fetal lipoproteins had a general pattern similar to that of adults, but high-density lipoproteins (HDL) were denser (1.128 g/ml) and correspondingly richer in proteins. The presence of apoprotein AI and of the light intestinal form of apoprotein B in the triglyceride-rich fractions as well as that of apoprotein AIV in HDL was indicative of an early contribution of enterocytes to the lipoprotein pool. Low-density lipoprotein apoproteins were heterogeneous until 14 days, as they are in the human fetus, and contained, besides apoprotein B, all the main apoproteins, HDL reached concentrations of 400 mg/100 ml at 3 and 4 weeks of age, above 3-fold higher than those of chow-fed adults, but also 1.5-fold higher than those of growing pigs fed a 20% fat diet, thereby showing a high capacity of the very young animal to synthesize HDL and apoprotein AI in particular. The many similarities with the evolution of lipoprotein in the newborn human validate the young pig as a model in the study of perinatal nutrition.

Synthesis and secretion of apolipoproteins by pig intestinal mucosa in organ culture: lack of inhibition of apolipoprotein A-I secretion by colchicine

Pig duodeno-jejunal mucosa was maintained in organ culture for up to 24 h in Eagles minimum essential medium containing 10% foal serum. Viability was controlled by determination of alkaline phosphatase and sucrase activity in the tissue. [14C]Leucine incorporation into proteins decreased 3-fold between 2 and 24 h. Newly synthesized secreted proteins were analyzed by SDS-polyacrylamide gel electrophoresis of the whole culture medium. Apolipoprotein A-I specifically measured by immunoelectrophoresis represented 10-20% of newly secreted proteins. Only 10% of apolipoprotein A-I secreted was recovered with the lipoprotein fraction (d less than 1.21). Recombination of the medium with porcine lipoproteins or DMPC vesicles prior to ultracentrifugation allowed, respectively, the recovery of 40 and 80% of apolipoprotein A-I secreted. The lipoprotein fractions also contained some apolipoproteins B and C and, after DMPC recombination, an apolipoprotein of Mr 45 000, most likely apolipoprotein A-IV, representing about 3.5% of newly secreted proteins. The d greater than 1.21 fractions all contained a high Mr protein, identified as IgA, and an unidentified protein of Mr approximately 45 000. The addition of colchicine (125 microM) to the culture medium did not significantly modify either tissue enzyme activities or [14C]leucine incorporation. It reduced total secretion by about 40% between 2 and 8 h of incubation, without interfering with apolipoprotein A-I secretion, which then represented up to 35% of secretion products. This raises the question of the mode of secretion of apolipoprotein A-I, which may be related to the high proportion of its which is secreted free.

Pig apolipoprotein AI self-association and interaction with L-α-dimyristoylphosphatidylcholine as compared with human apolipoprotein AI

Associative properties of apolipoprotein AI of HDL were compared in the pig and human. Self-association of apo AI (determined by dimethylsuberimidate cross-linking) occurred preferentially as tetramers in pig AI as compared to equal proportions of tetra and pentamers in human. Like human apo AI, the pig apoprotein is susceptible of recombining with increasing concentrations of phospholipids (dimyristoylphosphatidylcholine). Both apoproteins first give rise to particles containing 2 AI molecules (DMPC : AI molar ratio of 20 - 200) then to larger particles containing 3 apoprotein molecules (DMPC : AI molar ratio of 200 - 600). Despite these similarities in phospholipid association, pig apolipoprotein displayed less affinity for the lipid vesicles and should, therefore, be more readily exchangeable. Upon incubation with human HDL, however, pig apo AI displaces human apo AI, but it does not associate with it in mixed particles, and forms heavy particles from which the human apoprotein has been totally displaced. Protein-protein associations at the surface of lipoproteins therefore exhibit definite species specificity.